A sputum swab culture method for tubercle bacilli using Cetrimide, compared with two other swab culture methods and the concentration culture method.
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We have previously described a passive inflation method during constant inspiratory flow for measuring total respiratory elastance and resistance during mechanical ventilation in newborns. The simple method for measuring respiratory mechanics had been assessed during decelerating inspiratory flow obtained with pressure controlled ventilation (PCV). We report an application of this method to preterm and full-term mechanically ventilated newborn infants and a comparison with the occlusion method. Twenty-one newborn infants (birth weight 1,060 to 3,650 g; gestational age 26 to 41 weeks), between 1 to 55 days of postnatal age, were enrolled in the study. They were ventilated with a "Servo ventilator 900C," first set in the pressure-controlled mode and then in the volume-controlled mode without changing the tidal volume (VT), inspiratory time or ventilator rate. Flow was measured through a pneumotachograph inserted between the endotracheal tube (ETT) and the breathing circuit; VT was obtained by integration of flow and airway pressure measured directly at the airway opening. Flow, volume, and pressure were plotted on analog X/Y tables to obtain pressure-volume (P/V) and flow-volume (V/V) loops, as well as pressure-time curves. Occlusion was performed by using the end-inspiratory and the end-expiratory pause buttons of the ventilator. The passive inflation method during PCV was based on the analysis of P/V and V/V loops and provided compliance (Crs(PC)infl.), resistance Rrs(PC)infl.) of the respiratory system, and intrinsic positive end-expiratory pressure (PEEP(PC)i,infl.). These values were compared with (1) compliance (Crs(PC)occl.) and intrinsic positive end-expiratory pressure (PEEP(PC)i,occl.) measured by the occlusion method during PCV; (2) static (Crs(VC),occl.) and dynamic (Crs(VC),dyn.) compliance, airway (R(aw)(VC),), tissue (Rrs(VC),visc.) and total resistance (Rrs(VC),occl.), and intrinsic positive end-expiratory pressure (PEEP(VC)i,occl.) measured by the occlusion method during volume-controlled ventilation. Crs(PC),infl. correlated well with Crs(PC),occl., Crs(VC),occl., and Crs(VC),dyn.. Furthermore, Crs(PC),infl. and Crs(VC),dyn. did not differ significantly. Rrs(PC),infl. correlated well and did not differ significantly from total inspiratory resistance, i.e., the sum of R(aw)(VC) and Rrs(VC),vis. PEEPPC,i,infl. correlated well and did not differ significantly from PEEP(PC)i.occl. and from PEEP(VC),i,occl. The passive inflation method can be used during PCV with a decelerating flow waveform. It provides dynamic compliance, inspiratory resistance of the respiratory system, and intrinsic PEEP from the analysis of V/V and P/V loops recorded at the airway opening. This technique is simple to use and well tolerated by preterm and full-term ventilated newborn infants. It can be a good alternative to occlusion methods.
Two methods of dissecting rat fetuses are compared: the generally well-known freehand razor-blade dissection method according to Wilson and an in situ sectioning method (ISM). For this purpose the substance EGYT 1978 was used, which induces--inter alia--cardiovascular anomalies. Thirty-nine gravid rats were given either the test drug (EGYT 1978, 400 mg/kg body weight) or the vehicle orally from the 6th to the 15th day of gestation. The fetuses were assigned randomly to either dissection group. Both methods of examination revealed approximately the same number of anomalies per organ system. A comparison of the types of anomaly found showed, however, relevant differences in the nature of the changes revealed by the two methods. The in situ sectioning method was clearly superior to the freehand razor-blade dissection method for the detection of discrete anomalies of the cardiovascular system. We therefore recommend the in situ sectioning method as the method of choice for examining small fetuses from teratological experiments.
Fluorometric methods for determining delta-aminolevulinic acid (ALA) and coproporphyrin (CP) in urine have been recently developed by using high-performance liquid chromatography (HPLC). In the present study, urinary ALA and CP in lead-exposed workers were determined with these fluorometric HPLC methods and the conventional methods, and the results obtained were compared. In lead workers with a urinary ALA greater than or equal to 5 mg/l, the values obtained with the fluorometric HPLC method corresponded well with those measured with the conventional colorimetric method. In contrast, in lead workers with ALA less than 5 mg/l, ALA values obtained with the fluorometric HPLC method were lower than those measured with the conventional method, suggesting the possibility of matrix interference in urine. The urinary CP values obtained with the conventional method of Rimington (1971) were higher than those measured with the fluorometric HPLC method, though the correlation was good.
Until now methods using tetramethylbenzidine (TMB) for electron microscopy (TMB-EM methods) are all unable to provide a maximum demonstration of transported horseradish peroxidase (HRP) while maintaining good ultrastructural tissue preservation. In order to solve this problem, we have attempted to adapt a newly developed, highly sensitive TMB method using sodium tungstate (ST) as the stabilizer (TMB-ST method) for HRP electron microscopic retrograde and anterograde fiber tracing. The present study shows that the TMB-ST method combined with diaminobenzidine-cobalt (DAB-Co) is more sensitive than existing TMB-EM methods and that ultrastructural details are well preserved with this combined method. The resultant reaction product complex after osmication is stable and is observed as characteristic crystal-like structures which are extremely electron dense and often aggregated into clumps. In contrast, the TMB-ST method without the DAB-Co step frequently produces a moderate electron-dense reaction product. Therefore, we recommend the TMB-ST method combined with DAB-Co for HRP electron microscopy.
In the present investigation, the conventional drug susceptibility testing procedures using the 1% proportion method on Löwenstein-Jensen (LJ) and Middlebrook 7H10/7H11 agar media were compared with a recent radiometric method. The latter using "Bactec 460-TB" apparatus and a special broth medium (Becton and Dickinson Diagnostic Instrument Systems, Towson, MD, USA) gave drug susceptibility results within a week as compared to 4-5 weeks with the LJ method, and 3-4 weeks using 7H10/7H11 agar media. This report describes the critical concentrations retained for each of the methods and the methodology used. Also, the compatibility of the results obtained with the radiometric method with those obtained with conventional 7H11 agar method was perfect. In this report, we justify the choice of the 7H11 agar method instead of the LJ method as our conventional drug susceptibility procedure, and suggest also that radiometric drug susceptibility determination by Bactec 460-TB apparatus may be the method of choice for routine, rapid drug susceptibility determinations.
OBJECTIVE: To examine the qualitative and quantitative characteristics of frozen-thawed spermatozoa recovered through the SpermPrep (ZBL, Inc., Lexington, KY) filtration method or the swim-up technique for the purpose of intrauterine insemination (IUI) or other techniques for assisted reproduction. DESIGN, SETTING, PATIENTS: Thirty pairs of frozen specimens purchased from three commercial semen suppliers were used in this study. Each pair consisted of two aliquots from the same semen specimen. MAIN OUTCOME MEASURES: Spermatozoa recovered via the SpermPrep filtration and swim-up processes were evaluated for sperm numbers recovered, sperm motility, grade of sperm motility, percentage of morphologically normal spermatozoa, the response of spermatozoa to a hypo-osmotic environment (hypo-osmotic swelling test), and the amount of debris present. RESULTS: Application of the SpermPrep filtration method resulted in recovery of significantly greater numbers of spermatozoa (P less than 0.01) than were recovered with the swim-up method (31.1 +/- 3.2 x 10(6) versus 10.2 +/- 1.8 x 10(6) spermatozoa, respectively). This represents a mean recovery of approximately one half (49%) of all spermatozoa applied to the filter, whereas for the swim-up method, it was only 15%. The overall quality of recovered spermatozoa was virtually identical between the two methods (P greater than 0.05). The percent motile sperm improved by a mean of 18% to 20%, the grade of motility improved by a mean of 0.4 points (scale 0 to 4), the percent of morphologically normal spermatozoa increased by a mean of approximately 10%, the percent of spermatozoa reactive to a hypo-osmotic medium test increased by a mean of approximately 9%, and the debris score decreased by a mean of 0.2 to 0.3 points (scale 0 to 4). Most importantly, the mean total number of motile, morphologically normal spermatozoa after filtration through the SpermPrep column was 20.2 +/- 1.1 x 10(6), representing a mean recovery of 73% of the normal spermatozoa originally applied to the column. This was 316% greater than the yield obtained with the swim-up method (6.4 +/- 0.8 x 10(6)), which was significantly greater (P less than 0.01) than that recovered via the swim-up method. Also, the time required to harvest sperm through SpermPrep filtration was 20 to 25 minutes versus 80 minutes required for the swim-up method (P less than 0.05). CONCLUSION: Considering that the effectiveness of frozen-thawed semen is already limited when compared with fresh semen, SpermPrep filtration is the method of choice over the swim-up technique of sperm selection because the former provides significantly greater numbers of high quality sperm. It should be considered as an adjunct in semen preparation for IUI or other forms of assisted reproduction.
The authors have developed a new scanning method for Tl-201 myocardial SPECT using a rotating gamma camera with 180-degree data collection. This requires a new type of imaging table on which the patient is placed in a 50-degree right semidecubital position to allow close proximity to the camera-collimator system. Using this method, attenuation by the imaging table is completely avoidable because the projected views in the left oblique directions are not obstructed by the table. In addition, the radius of camera rotation is about 25% shorter than in the conventional method. Phantom and 16 clinical studies were performed to compare the performance of conventional and semidecubital position methods. In phantom studies, tomographic images produced using the new method showed better lesion contrast and more counts in the posteroinferior wall of the heart than those produced using the conventional method. Clinically, 12 of the 16 studies showed more counts in the posteroinferior wall of the heart with the semidecubital position method than with the conventional method. The semidecubital position method is useful for Tl-201 myocardial SPECT, especially for evaluation of posteroinferior wall hypoperfusion.
A study to compare three different methods for reading MIC endpoints tested by the broth microdilution modification of the National Committee for Clinical Laboratory Standards (Villanova, Pa.) reference method was conducted. MICs of amphotericin B, flucytosine, fluconazole, itraconazole, and a new triazole, D0870, were determined for five reference yeast strains and 100 clinical isolates of Candida spp. MICs were read visually according to National Committee for Clinical Laboratory Standards guidelines from microdilution trays that had been (VS) and had not been (V) shaken. MICs were also determined spectrophotometrically (SP) at 492 nm. SP endpoints were determined as the concentrations resulting in a > or = 50% inhibition of growth (flucytosine and azoles) and a > or = 90% inhibition of growth (amphotericin B) relative to control growth. The five reference strains were tested nine times each against all five antifungal agents, and the MIC results for each reading method were compared with a 3-log2 dilution reference range determined by the macrodilution (M27-P) method. Overall, 84 to 100% of the MICs determined by V, 93 to 100% of those determined by VS, and 89 to 100% of those determined by SP fell within the 3-log2 dilution reference range for each reference strain and antifungal agent. Reproducibility was 99% for V and SP and 98% for VS. Agreement among the three methods of reading ranged from 97 to 99%. Excellent agreement among reading methods was also observed for all antifungal agents when tested against 100 clinical isolates. Agreement between the standard V method (no agitation) and VS ranged from 99 to 100%, and that between V and SP ranged from 89 to 99%. The VS and SP reading methods provided more definitive endpoints than the V method, which does not involve shaking.
We report studies of the validity and clinical application of a new amidolytic method for evaluating the activated partial thromboplastin time (APTT) compared with a conventional clotting method. The results with the two methods were well correlated for normal plasma and plasma from hemophilia patients. Congenital deficiencies of of the intrinsic coagulation pathway other than hypo- and dysfibrinogenemia detected by the amidolytic method agreed with those detected by the clotting APTT. The results with the two methods for plasma from patients under heparin treatment were statistically different, suggesting a lesser sensitivity of the amidolytic method to heparinization. The use of the amidolytic APTT reagent in combination with factor VIII and IX deficient plasma allowed the measurement of both factors. The results obtained with normal and hemophilic plasma were in agreement with those obtained with the one-stage clotting method in all except two occasions. Even though the amidolytic method demonstrated the presence of the lupus anticoagulant in the majority of tested samples of known lupus subjects, it was less sensitive to the abnormality than the clotting method.
We describe a "high-performance" liquid chromatographic (HPLC) method for accurately determining creatinine in serum. After prechromatographic precipitation of protein, we performed isocratic ion-exchange chromatography with ultraviolet detection (234 nm). Analytical results showed linearity up to 1770 mumol/L, a detection limit of 22 mumol/L, an average analytical recovery of 101%, and a CV ranging from 3% to 11%. We used certified human serum (National Institute of Standards and Technology), and additional lyophilized serum pools also assayed by definitive isotope-dilution mass spectrometry, to validate the accuracy of the HPLC method. In addition, the isocratic HPLC results showed close agreement with those obtained with a step-gradient HPLC method. We also compared the isocratic HPLC method with alkaline picrate and enzymatic methods. Our findings with samples from nonuremic, uremic, and diabetic ketoacidotic patients confirmed the positive bias previously reported with the alkaline picrate method. Interlaboratory transferability of the method was demonstrated with various commercial instruments and analytical columns. We evaluated column stability and possible interference from endogenous or exogenous compounds. On the basis of our analytical findings, we recommend the isocratic HPLC method as a candidate Reference Method for determining creatinine in serum.
Three liquid chromatographic (LC) methods employing amperometric detection were compared with the collaboratively studied FDA optimized Monier-Williams distillation method for the determination of total sulfite in 5 food types. The foods included lemon juice, white wine, instant mashed potatoes, golden raisins, and onion flakes. Two of the LC methods (one employing headspace sampling and the other direct injection) used ion-exchange chromatography with a basic mobile phase (pH about 10.8) and a glassy carbon electrode; the third (employing direct injection) used ion-exclusion chromatography with an acidic mobile phase (pH about 2) and a platinum electrode. All 4 methods produced similar results for the wine, lemon juice, and raisins. Results were different for instant mashed potatoes and onion flakes. The headspace-LC method and direct ion-exclusion LC method, both of which employed an alkaline sample extraction, yielded significantly higher values for sulfite in instant potatoes than did the other 2 methods. A large interfering peak with both direct LC methods prevented quantitation of sulfite in the onion flakes. All methods can detect sulfite as low as about 1 microgram/g in 4 of 5 food types examined.
In order to evaluate the usefulness of various quantitative methods for the assessment of regional left ventricular function, a total of 9 methods (3 hemiaxial models, 1 area model and 5 radial-axes models) were applied to the radiopaque ventriculograms of 16 patients with anterior wall infarction and 13 patients with posterior wall infarction. A series of 17 patients without clinical or angiographic evidence of heart disease were used as control group. While the hemiaxial methods were consistently found to be useful and reproducible, the area method did not provide significant additional information. The diagnostic accuracy was highest with the use of 2 radial-axes methods. The sensitivity and specificity data were 90% to 100% and 70% to 100%, respectively. The remaining 3 radial-axes methods were found to be of limited value since regional wall motion abnormalities were projected to the contralateral wall. Because of this methodological shortcoming these techniques may falsely indicate asynergy in areas with normal contraction patterns. This was reflected by high sensitivity but low specificity values. A feature of the latter 3 methods was the use of the gravity center of the end-systolic frame as the reference point. The position of this reference point is decisively influenced by the extent and localization of asynergy. These findings suggest that methods using the gravity center of the end-systolic frame should not be employed in the evaluation of regional left ventricular function. In general, some quantitative methods for assessing regional left ventricular function require a considerable expense for technical apparatus which is not always justified by the extent to which additional information is gained.